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The Effects of Fenarimol and Methyl Parathion on Glucose 6-Phosphate Dehydrogenase Enzyme Activity in Rats

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The Effects of Fenarimol and Methyl Parathion on Glucose 6-Phosphate Dehydrogenase Enzyme Activity in Rats Türkiye Tarımsal Araştırmalar Dergisi Turk J Agric Res 2017, 4(3): 275-280 dergipark.gov.tr/tutad © TÜTAD ISSN: 2148-2306 e-ISSN: 2528-858X Research Article doi: 10.19159/tutad.310371 The Effects of Fenarimol and Methyl Parathion on Glucose 6-Phosphate Dehydrogenase Enzyme Activity in Rats Ferda ARI*, Egemen DERE, Hakan TOSUNOĞLU, İmren ALİOĞLU Uludağ University, Faculty of Science and Art, Department of Biology, Bursa, TURKEY Received: 03.05.2017 Accepted: 26.08.2017 ORCID ID (By author order) ORCID ID (by author order) İD orcid.org/0000-0002-6729-7908 İD orcid.org/0000-0001-9572-1051 İD orcid.org/0000-0001-7878-3373 İD orcid.org/0000-0003-0687-9362 orcid.org/0000-0002-6729-7908; orcid.org/0000-0001-9572-1051; orcid.org/0000-0001-7878-3373; orcid.org/0000-0003-0687-9362 *Corresponding Author: [email protected] *Corresponding Author: [email protected] Abstract: Fenarimol and methyl parathion are pesticides that have been used in agriculture for several years. These pesticides have significant effects on environmental and human health. Therefore, we investigated the effects of methyl parathion and fenarimol on glucose 6-phosphate dehydrogenase (EC 1.1.1.49) enzyme activity in rats. The glucose 6- phosphate dehydrogenase is the first enzyme of the pentose phosphate pathway and it is important in detoxifying reactions by NADPH generated. In this study, wistar albino rats administrated with methyl parathion (7 mg kg–1) and fenarimol (200 mg kg−1) by intraperitoneally for different periods (2, 4, 8, 16, 32, 64, and 72 h). The glucose 6-phosphate dehydrogenase enzyme activity was assayed in liver, kidney, brain, and small intestine in male and female rats. The exposure of fenarimol and methyl parathion caused increase of glucose 6-phosphate dehydrogenase enzyme activity in rat tissues, especially at last periods. We suggest that this increment of enzyme activity may be the reason of toxic effects of fenarimol and methyl parathion. Keywords: Glucose 6-phosphate dehydrogenase, fenarimol, methyl parathion, pesticide 1. Introduction pathways including specific enzymes. Glucose-6- phosphate dehydrogenase is the first enzyme on Pesticides are chemical substances, designed for pentose phosphate pathway (Frederiks and eliminating, preventing, or controlling variety of Vreeling-Sindelárová, 2001; Beydemir et al., pests. Pesticides have subgroups such as 2003). This enzyme is able to produce ribose 5- insecticides, fungicides, acaricides, herbicides phosphate and NADPH which are essential cellular according to targets, activity, and structure (Arias- systems on antioxidant pathway, membrane lipids Estévez et al., 2008). Generally they used mixtures synthesis, cytochrome p450 system reductive and and have potential adverse effects on human health nucleic acid synthesis (Stanton, 2012). and ecosystem (Karadeniz and Yenisoy-Karakaş, 2015). Fenarimol is a pyrimidine-type fungicide In this study, we investigated the effects of used in agriculture, especially for protection from fenarimol and MP on G6PD enzyme activity in the fungal spores or fungi (Paolini et al., 1996; Zhang liver, kidney, brain and small intestine tissues in et al., 2012; Oh et al., 2015). Methyl parathion rats and according to our investigation, there is no (MP) is a widely used organophosphate information in literature about the effects of insecticides in agriculture that is used to kill fenarimol and MP on G6PD enzyme activity in rat insects on crops (Anonymous, 1998). Fenarimol tissues. The present study will provide further and MP have potential toxic effects on organisms insight on toxic mechanism in tissues of rats and environment due to widespread usage. They exposed to fenarimol and MP. also affected living organism and metabolic ARI et al. 2. Materials and Methods by activation or inhibition (Beydemir et al., 2003; Ekinci and Beydemir, 2010). In the literature, there 2.1. Animals are various studies about the effects of pesticides Wistar albino rats (Rattus norvegicus) divided on G6PD activity (Kaur and Sandhu, 2008; into control (2 male; 2 female) and experimental Şentürk et al., 2009; Ojha and Srivastava, 2012; groups (4 male; 4 female). All animals (200-250 g) Salvo et al., 2012; Topal et al., 2014). However, were purchased from Experimental Animals there is no data about G6PD responses of Feeding and Research Centre of Uludağ fenarimol and MP on rat tissues. Thus, it is University. Rats housed in specialized animal important to obtain information about the effects room on 12-h light/dark cycle at 21-23 oC and they of fenarimol and MP on rats. were treated with corn oil while experimental groups were injected intraperitoneally with 200 mg Glucose 6-Phosphate Dehydrogenase is known -1 as a useful biomarker for antioxidant system and kg (LD50) dose of fenarimol (Sigma-Aldrich, St. -1 its activity is increased in oxidative stress Louis, MO) and 7 mg kg (LD50) dose of MP (Sigma-Aldrich, St. Louis, MO). The rats were left (Salvemini et al., 1999; Gül et al., 2004). In without food and water for 24h prior to injection, present study, we investigated the effects of ensuring the simultaneous initiation of metabolism fenarimol and MP on G6PD activities in some of animals in both groups at the same time and tissues. The effect of fenarimol and MP on liver after the injection phase, food and water were G6PD activity was determined in male and female regularly given to the animals until the trial rats. The G6PD activity did not change after periods were completed. Treated and control rats treatment of fenarimol and MP in liver at first were kept in plastic metabolic cages. Animals were hours in experimental periods (2, 4 and 8 hours) euthanized via cervical dislocation at each time while the G6PD activity generally increased at point (2, 4, 8, 16, 32, 64, and 72h post-injection). later periods (16, 32 and 64 hours) in rats. Similarly, significant difference at 72 hours was detected in male rats by fenarimol (p<0.05). 2.2. Determination of glucose 6-phosphate However, the G6PD activation at 32 and 64 hours dehydrogenase (G6PD) activity are significant in female rats by fenarimol (p<0.05) The liver, kidneys, brain and small intestine (Figure 1). were quickly removed and were perfused in ice- cold 0.15 M KCl, immediately after the In kidney, the G6PD activity significantly homogenates were prepared and homogenized at increased by fenarimol at 16, 32, 64 and 72 hours 2000 rpm in a T-line laboratory stirrer type in male and female rats compared with control homogenizer. Each homogenate was centrifuged in group (p<0.05). These activations reached a Dupont Instruments Sorvall “RC-5 super speed maximum level at 16th hour and decreased through refrigerated centrifuge” at 48000 g for 30 minutes. the end of the experimental periods (Figure 2a and Protein concentration was determined with the 2b). However, the G6PD activity did not method of Bradford (Bradford, 1976) and bovine significantly change by methyl parathion in kidney serum albumin was used as protein standard. The tissues of rats (Figure 2c and 2d). glucose 6-phosphate dehydrogenase activity was It is found that G6PD activity is increased in assessed spectrophotometrically via the Bohringer liver, and kidneys treated with fenarimol and MP, Manheim method (Bohringer, 1973). especially in male rats. These increments may be responsible to get rid of the toxic effects of MP 2.3. Statistical analysis and fenarimol, since G6PD plays an important role Data were analyzed using SPSS 20.0 for windows in detoxification pathway. Similarly, Rodriguez- and independent t-test was applied between data of Ariza et al. (1993) found that G6PD activity was control and experiment periods. The significance increased after pesticide toxicity in gray mullet was calculated using one-way analysis of variance (Mugil sp.). In another study, it was shown that (ANOVA) and Student’s t-test and as a result a G6PD activity was increased in gill of fresh water value of p<0.05 was taken as statistically mussel, Lamellidens marginalis by MP (Moorthy significant. et al., 1985). Effects of fenarimol and MP on G6PD activity 3. Results and Discussion in brain tissues of rats are shown in Figure 3. Pesticides are potential chemical pollutants which According to the results, fenarimol exhibited have increasing usage in agriculture and they have increased G6PD activity at last experimental harmful effects on the environment and organisms periods (8, 16, 32 and 64 hours) compared with the (Igbedioh, 1991; Aktar et al., 2009). Pesticides controls in male and female rats. These increases change metabolism, especially enzyme activities are significant (p<0.05) at 8, 32 and 64 hours in 276 T ü r k i y e T a r ı m s a l A r a ş t ı r m a l a r D e r g i s i - T u r k i s h J o u r n a l o f A g r i c u l t u r a l R e s e a r c h 4 ( 3 ) : 2 7 5 - 2 8 0 ARI et al. Figure 1. Effects of fenarimol (a, b) and methyl-parathion (c, d) on glucose 6-phosphate dehydrogenase (G6PD) activity (U/mg protein) in liver of rats *: Denotes statically significant differences in comparison with control (p<0.05) Figure 2. Effects of fenarimol (a, b) and methyl-parathion (c, d) on glucose 6-phosphate dehydrogenase (G6PD) activity (U/mg protein) in kidney of rats *: Denotes statically significant differences in comparison with control (p<0.05) Türkiye Tarımsal Araştırmalar Dergisi - Turkish Journal of Agricultural Research 4(3): 275-280 277 ARI et al. brain tissues of female rats (Figure 3a and 3b). metabolized in the liver and ejaculated through the Although, the G6PD activity was decreased at 16 kidney, so we found the G6PD activity was higher and 32 hours in male rats by methyl parathion in liver and kidney through fenarimol and MP (Figure 3c and 3d).
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